Multispectral imaging for MicroChip electrophoresis enables point-of-care newborn hemoglobin variant screening

An, Ran; Huang, Yu‐Ning; Rocheleau, Anne D.; Avanaki, Alireza; Thota, Priyaleela; Zhang, Qiaochu; Man, Yuncheng; Sekyonda, Zoe; Segbefia, Catherine; Dei‐Adomakoh, Yvonne; Mensah, Enoch; Ohene‐Frempong, Kwaku; Odame, Isaac; Owusu-Ansah, Amma; Gürkan, Umut A.

doi:10.1016/j.heliyon.2022.e11778

Cited by 10 publications

(6 citation statements)

References 47 publications

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“…The Gazelle TM system design, including the microcartridge structure, paper-based electrophoresis principle, instrument design, and optical path structure, can be found in our previous publications [30,32]. The technicians performed the tests according to the Gazelle-Multispectral instructions for use as published previously [30].…”

Section: Gazelle Test Proceduresmentioning

confidence: 99%

“…The fundamental principle behind Gazelle TM is hemoglobin electrophoresis in which different hemoglobin variants can be separated based on mobility differences under an electric field (Figure 1D). Gazelle has been tested in clinical studies in four different countries with more than 700 subjects and demonstrated a capability of identifying major Hb variants, including HbA, HbE, HbS, and HbF, in adults as well as in newborns with SCD, sickle cell trait, hemoglobin C disorder, and hemoglobin E disorder [28,[30][31][32].…”

Section: Introductionmentioning

confidence: 99%

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Point-of-Care Diagnostic Test for Beta-Thalassemia

An,

Avanaki,

Thota

et al. 2024

Biosensors

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Hemoglobin (Hb) disorders are among the most common monogenic diseases affecting nearly 7% of the world population. Among various Hb disorders, approximately 1.5% of the world population carries β-thalassemia (β-Thal), affecting 40,000 newborns every year. Early screening and a timely diagnosis are essential for β-thalassemia patients for the prevention and management of later clinical complications. However, in Africa, Southern Europe, the Middle East, and Southeast Asia, where β-thalassemia is most prevalent, the diagnosis and screening for β-thalassemia are still challenging due to the cost and logistical burden of laboratory diagnostic tests. Here, we present Gazelle, which is a paper-based microchip electrophoresis platform that enables the first point-of-care diagnostic test for β-thalassemia. We evaluated the accuracy of Gazelle for the β-Thal screening across 372 subjects in the age range of 4–63 years at Apple Diagnostics lab in Mumbai, India. Additionally, 30 blood samples were prepared to mimic β-Thal intermediate and β-Thal major samples. Gazelle-detected levels of Hb A, Hb F, and Hb A2 demonstrated high levels of correlation with the results reported through laboratory gold standard high-performance liquid chromatography (HPLC), yielding a Pearson correlation coefficient = 0.99. This ability to obtain rapid and accurate results suggests that Gazelle may be suitable for the large-scale screening and diagnosis of β-Thal.

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Section: Gazelle Test Proceduresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Point-of-Care Diagnostic Test for Beta-Thalassemia

An,

Avanaki,

Thota

et al. 2024

Biosensors

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“…Instrumental platforms typically used for such purposes as mentioned in the introduction (e.g., CE, HPLC, or MS/MS) are usually complex and costly. However, in this respect, qPCR-based primary screening for HbS alleles and thus reducing the sample size to a group of specimens relevant to further investigations might be a promising approach to open SCD screening to less complicated and/or inexpensive methods, such as chip-based microelectrophoresis, aqueous multi-phase systems based on cell density measurements, lateral flow immunoassays, or even classical electrophoresis [23,[36][37][38].…”

Section: Plos Onementioning

confidence: 99%

A high-throughput newborn screening approach for SCID, SMA, and SCD combining multiplex qPCR and tandem mass spectrometry

Tesorero

Janda²,

Hörster³

et al. 2023

PLoS ONE

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Early diagnosis of severe combined immunodeficiency (SCID), spinal muscular atrophy (SMA), and sickle cell disease (SCD) improves health outcomes by providing a specific treatment before the onset of symptoms. A high-throughput nucleic acid-based method in newborn screening (NBS) has been shown to be fast and cost-effective in the early detection of these diseases. Screening for SCD has been included in Germany’s NBS Program since Fall 2021 and typically requires high-throughput NBS laboratories to adopt analytical platforms that are demanding in terms of instrumentation and personnel. Thus, we developed a combined approach applying a multiplexed quantitative real-time PCR (qPCR) assay for simultaneous SCID, SMA, and 1st-tier SCD screening, followed by a tandem mass spectrometry (MS/MS) assay for 2nd-tier SCD screening. DNA is extracted from a 3.2-mm dried blood spot from which we simultaneously quantify T-cell receptor excision circles for SCID screening, identify the homozygous SMN1 exon 7 deletion for SMA screening, and determine the integrity of the DNA extraction through the quantification of a housekeeping gene. In our two-tier SCD screening strategy, our multiplex qPCR identifies samples carrying the HBB: c.20A>T allele that is coding for sickle cell hemoglobin (HbS). Subsequently, the 2nd tier MS/MS assay is used to distinguish heterozygous HbS/A carriers from samples of patients with homozygous or compound heterozygous SCD. Between July 2021 and March 2022, 96,015 samples were screened by applying the newly implemented assay. The screening revealed two positive SCID cases, while 14 newborns with SMA were detected. Concurrently, the qPCR assay registered HbS in 431 samples which were submitted to 2nd-tier SCD screening, resulting in 17 HbS/S, five HbS/C, and two HbS/β thalassemia patients. The results of our quadruplex qPCR assay demonstrate a cost-effective and fast approach for a combined screening of three diseases that benefit from nucleic-acid based methods in high-throughput NBS laboratories.

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“…Sickle cell disease (SCD) is a serious disease leading to circulatory disorders, organ damage, severe pain and early death [1]. According to current figures, about 400,000 are born with SCD every year [2]. The term SCD comprises a group of hemoglobin (Hb) disorders, which are autosomal recessively inherited and characterized by the presence of hemoglobin S (HbS) resulting from a point mutation in the 6 th codon of the β-globin encoding HBB gene (HBB CD 6 GAG > GTG [Glu > Val]).…”

Section: Introductionmentioning

confidence: 99%

High Throughput Newborn Screening for Sickle Cell Disease – Application of Two-Tiered Testing with a qPCR-Based Primary screen

Janda,

Hegert,

Bzdok

et al. 2023

Klin Padiatr

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Background Sickle cell disease (SCD) is a group of hemoglobinopathies with a common point mutation causing the production of sickle cell hemoglobin (HbS). In high-throughput newborn screening (NBS) for SCD, a two-step procedure is suitable, in which qPCR first pre-selects relevant samples that are differentiated by a second method. Methods Three NBS centers using qPCR-based primary screening for SCD performed a laboratory comparison. Methods using tandem MS or HPLC were used for differentiation. Results In a benchmarking test, 450 dried blood samples were analyzed. Samples containing HbS were detected as reliably by qPCR as by methods established for hemoglobinopathy testing. In a two-step screening approach, the 2nd-tier-analyses have to distinguish the carrier status from pathological variants. In nine months of regular screening, a total of 353,219 samples were analyzed using two-stage NBS procedures. The 1st-tier screening by qPCR reduced the number of samples for subsequent differentiation by>99.5%. Cases with carrier status or other variants were identified as inconspicuous while 78 cases with SCD were revealed. The derived incidence of 1:4,773, is in good agreement with previously published incidences. Conclusion In high-throughput NBS for SCD, qPCR is suitable to focus 2nd-tier analyses on samples containing HbS, while being unaffected by factors such as prematurity or transfusions. The substantial reduction of samples numbers positively impacts resource conservation, sustainability, and cost-effectiveness. No false negative cases came to attention.

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Multispectral imaging for MicroChip electrophoresis enables point-of-care newborn hemoglobin variant screening

Cited by 10 publications

References 47 publications

Point-of-Care Diagnostic Test for Beta-Thalassemia

Point-of-Care Diagnostic Test for Beta-Thalassemia

A high-throughput newborn screening approach for SCID, SMA, and SCD combining multiplex qPCR and tandem mass spectrometry

High Throughput Newborn Screening for Sickle Cell Disease – Application of Two-Tiered Testing with a qPCR-Based Primary screen

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